Purification of immunoglobulins

Abstract

The present invention relates to a separation matrix comprised of porous particles to which antibody-binding protein ligands have been immobilised, wherein the ligand density is in the range of 5.0-10 mg / ml; the gel phase distribution coefficient of the particles expressed as Kav for a dextran of size 110 kDa is above 0.65 and the median particle diameter is between 65-84 μm. The carbohydrate material is preferably highly cross-linked agarose.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims priority to U.S. provisional patent application number 60 / 638,316 filed Dec. 22, 2004; the entire disclosure of which is incorporated herein by reference in its entirety.FIELD OF THE INVENTION [0002] The present invention relates to the field of antibody preparation, and more specifically to a separation matrix for isolation and / or separation of antibodies. The invention also encompasses a chromatography column that comprises the separation matrix of the invention, a method of isolating antibodies using said separation matrix and a multistep process for large-scale purification of antibodies from a crude feed. BACKGROUND OF THE INVENTION [0003] The immune system is composed of many interdependent cell types that collectively protect the body from bacterial, parasitic, fungal, viral infections and from the growth of tumour cells. The guards of the immune system are macrophages that continually roam the bloodstream...

AI Technical Summary

Benefits of technology

[0014] One aspect of the present invention is a novel separation matrix suitable for purification of polyclonal or monoclonal antibodies. This can be achieved as defined in the claims. Another aspect of the invention is a separation matrix which allows faster and more economic purification of antibodies than the prior art. This can be achieved by a novel separation matrix as defined in the appended claims, which enables a substantially increased binding capacity when used in chromatography.

Problems solved by technology

Typically, these precipitation methods are giving very impure products while at the same time being time consuming and laborious.

Furthermore, the addition of the precipitating agent to the raw material makes it difficult to use the supernatant for other purposes and creates a disposal problem, which is particularly relevant when speaking of large-scale purification of immunoglobulins.

Thus, a disadvantage of this procedure is the necessity to add lyotropic salt to the raw material as this gives and problem and thereby increased cost to the large-scale user.

For other raw materials than cell culture supernatants such as whey, plasma, and egg yolk the addition of lyotropic salts to the raw materials would in many instances be prohibitive in large-scale applications as the salt could prevent any economically feasible use of the immunoglobulin depleted raw material.

An additional problem in large-scale applications would be the disposal of several thousand litres of waste.

Although the matrices described for thiophilic chromatography generally show good performance, since lyotropic salts are added to the raw material to ensure efficient binding of the immunoglobulin, disadvantages as discussed above will arise.

However, despite the state of the art constructions, there is still a need of alternative separation matrices for purification of antibodies or antibody constructs, which observe the demands of purity, safety, potency and cost effectiveness.

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